Apparatus for chromatographic analysis



June 8, 1965 J. HRDINA 3,187,786

APPARATUS FOR CHROMATOGRAPHIC ANALYSIS Filed March 3, 1964 2 Sheets-Sheet l Zjvwwkw J HRDINA June 8, 1965 J. HRDlNA 3,187,786

APPARATUS FOR CHROMATOGRAPHIC ANALYSIS Filed March 3, 1964 2 Sheets-Sheet 2 FIG. 3

grave/114M J|R| HRDINA 3,187,786 APPARATUS FOR CHROMATGGRAPIHC ANALYSIS Jiri Hrdina, Prague, Czechoslovakia, assignor to Ceslroslovenska akademie ved, Prague, Czechoslovakia, a corporation of Czechoslovakia Filed Mar. 3, 1964, Ser. No. 349,055 Claims priority, application Cz/e6choslovakia, Sept. 3, 1960,

i 4 Claims. (Cl. 141-94) This application is a continuation-in-part of my copending application Serial No. 135,542 filed on September l, 1961 (now abandoned), claiming the filing date of my Czechoslovak patent application of September 3, 1960, of which a certified copy has been placed of record inthe file.

My invention relates to an apparatus for controlling the steps in methods for the resolution of mixtures and for the isolation and identification of the components,

known as chromatography or chromatographic analysis.

In such processes using columns for the mobile phase, which may be either liquid or gaseous, the fractions of eluate leaving the columns are collected in individual containers, which have to be consecutively exchanged either manually or automatically. The time intervals for such exchanges are generally either quite freely assessed or guessed without regard to the actual progress of the chromatographic process, or they are elected in dependence of the amount of the mobile phase passed through the column; or in dependence of the precedingly elected weight or volume of the collected eluate.

The said known methods are quite complicated and inexact because the relation between the processual determinants is unforseeable. Thus for example the premise is that the collection of eluate in a certain period of tirne is commensurate with the collection measured by volume, does not apply when changes or" the hydraulic conditions occur in view of any swelling of the content of the columns which causes changes of the velocity of the flow. 'Often the chromatographic process must be undesirably interrupted when the pump delivering the mobile phase into'the columns causes such rise of pres sure that the columns are endangered.

The said and other disadvantages are avoided by my invention. The invention provides an apparatus for use as a fraction collector in chromatographic analysis wherein, according to practices common in the art of chromatography, a liquid or gas (mobile phase) is uninterruptedly passed through or in contact with a column of adsorbent or separation substance (solid phase) which is contained in a tube and wherein the resutling eluate issues from the outlet of the tube drop by drop. In accordance therewith I use a volumetrically precise piston pump which delivers exactly measured volumes of the mobile phase to the columns and cooperatively couple the pump with an electromechanical device which controls the exchange of containers for succeeding fractions so that the exchange of containers is initiated or effected only in dependence on the number of strokes of the piston of the precision pump without regard whether this pump is slowly or quickly or uniformly driven and without regard to the number and the unpredictable size of the droplets of eluate leaving the column.

The beforesaid and other objects of my invention will be more fully understood from the following specification when read with the accompanying drawing illustrating embodiments of my invention.

In the drawing FIGJ illustrates schematically a chromatographic apparatus with manual exchange of the receiving vessels,

FIG. 2 illustrates schematically a chromatographic ap- United States Patent 0 "ice 3,187,786 Patented June '8, 1965 paratus with automatic exchange of the receiving vessels, and

FIG. 3 shows an elevational view partly in sectiono't a constructively developed embodiment of my chromatographic apparatus.

The same reference characters indicate the same or equivalent elements in all figures.

The embodiment shown schematically in FIG. 1 comprises a precision piston pump 12 which delivers volumetrically identical fractions of the mobile phase through the pipeline 13 into the chromatographic column 14 which includes an adsorbent or separation substance (solid phase) which is contained within a vertically disposed discharge tube 13. Each of said fractions of 'the mobile phase which is volumetrically proportional to the number 'of strokes of the piston of the .pump, passes as known through the column 14 and out via the outlet of discharge tube 13' and is collected in a flask or other vessel 15 which when filled is replaced by an empty vessel to receive next following fractions.

Accoustical or optical signaling means, for example an electrically operated clock 16, indicate the time when an exchange of the vessel 15 has to be made. The said alarm means 16 is operated over the electrical conduits 18 by an electromechanical device 17 which is operatively connected to the pump 12 by the rod 19. Thus -the"signa'ling means 16 is actuated after a predetermined number of strokes of the piston of pump 12 and the deliveryof apredetermined volume of the mobile phase into the column 14.

The embodiment schematically shown in FIG. 2 includes like that shown in FIG. 1 a precisionpump 12and a pipeline 13 leading into the chromatographic column 14. A plurality of equally spaced vessels such as retorts 36 is placed in one of the known fraction collecting apparatus 37. This apparatus is stepwise operated by the electromechanical device 17, to which it' is electrically connected by the conduits 18, and it places one retort 36 after the other under the outlet of tube 13 in which column 14 is contained. The electromechanical device 17 is actuated in the same manner as in the embodiment illustrated in FIG. 1. Thus in both embodiments the precision pump 12 not only delivers predetermined volumes of the mobile phase into the column 14 but also controls the required exchange of the collecting vessels 15 and 37, respectively.

FIG. 3 shows a constructively developed embodiment of my invention. The precision pump 12 driven by the electric motor 20 over the worm 21 and wormwheel 22 sucks the mobile phase throughpipeline 11 and delivers it through pipeline 13 into the vertically disposed chromatographic column 14 which may be held in a standard 14'. The eluate leaving the column 14 and issuing from the outlet of tube 13' is collected in exchangeable flasks 15 or in retorts .36 mounted in a rotatable fraction-colleo tion apparatus 35, only part of it being shown.

An electromechanical device 23 to 29 is operated by the pump 12 by means of the connecting rod 22' and actuates an optical or accoustical signaling device 16 to indicate the time for removal of a filled flask 15, or con trols the motor 33 which drives over the worm gear 37 the fraction-collecting apparatus 35 with retorts 36.

The said electromechanical device includes a worm 23 driven by the wheel 22 and driving a changeable gearing 24, 25 and 26 which is operatively coupled with an exchangeable cam 27. This cam actuates a switch 28 in a circuit 30 activated by the battery 29.

The circuit 30 operates either the bell .16 or over the relay 31 the switch 32 which connects the motor 33 to an electrical network or other power source 38. The motor 33 drives over the gearing 37 the rotatable fracdon-collecting apparatus 35 which stepwise and synchrolets.

nously with a predetermined number of strokes of the piston of the pump 12 places one retort 36 after the other under the column 14.

It will be well understood that the apparatus according to my invention may be used for such other purposes than chromatographic analysis, wherein the processing of a liquid is made directly dependent on the volume of the supplied liquid. My invention avoids complicated and dangerously sensitive means which have inexact and uncontrollable efiects. Typical in this respect are the known apparatus wherein the amount of liquid entering the collecting vessel is measured by the numbers of drop- Such measurement is unreliable and inexact be cause it is dependent on the unpredictable surface tension of the liquid, the unpredictable size of the droplets, and of course on the unknown composition of the liquid to 'be analysed.

Though specific embodiments of my invention have been shown and described in detail to illustrate the application of the principles of my invention, it will be understood that the same may be otherwise embodied without departing from such principles.

- What I claim as my invention is: Y

1. In an apparatus for chromatographic analysis of a mobile phase and for collection of resulting fractions of eluate, the improvement which comprises in combination:

a substantially vertically disposed column including a tube and a separation substance;

a precision pump delivering a mobile phase to an upper part of said column;

a piston in said pump and a first driving means reciprocating the piston;

an exchangeable collection vessel adapted to receive a fraction of eluate issuing from a lower part of said column;

signaling means indicating a desired replacement of said collecting vessel by another collecting vessel. to receive a further fraction of eluate;

electromechanical means operating the signaling means in synchronous relation to a predetermined succession of strokes of the pump piston;

and second driving means for said electromechanical meansoperatively connecting said electromechanical means with said first driving means.

2. Apparatus for chromatographic analysis. and for collection of fractions of eluate according to claim 1, in which said electromechanical means includes a gearing driven by the reciprocating pump piston;

an exchangeable cam driven by said gearing;

, 4 and an electrical switch controlling the signaling means and operated by said cam in synchronous relation to a predetermined succession of strokes of the pump piston.

3. In an apparatus for chromatographic analysis of a mobile phase and for collection of resulting fractions of eluate, the improvement which comprises in combination:

a substantially vertically disposed column including a tube and a separation substance;

a precision pump delivering a mobile phase to an upper part of said column;

a piston in said pump and first driving means reciproeating the piston;

a fraction collecting device including a plurality of collecting vessels to be successively placed underthe column and adapted to receive succeeding fractions of eluate issuing therefrom;

an electric motor operating said fraction collecting device;

electromechanical means controlling the operation of said motor in synchronous relation to a predeten' References Cited by theExaminer UNITED STATES PATENTS 2,604,249 7/52 Gorham 141-130 2,660,336 11/53 Coffey et al. 222-3'72 XR 2,672,581 3/54 Gorham 141-130 2,870,941 1/59 Steidley 222- 2,880,764 4/59 Pelavin 141-130 3,018,922 1/ 62 Williamson 22270 XR 3,074,445 1/ 63 Rosen ,222-70 XR 3,124,172 3/64 Paxson 141-430 LAVERNE D. GEIGER, Primary Examiner. 

1. IN AN APPARATUS FOR CHROMATOGRAPHIC ANALYSIS OF A MOBILE PHASE MAND FOR COLLECTION OF RESULTING FRACTIONS OF ELUATE, THE IMPROVEMENT WHICH COMPRISES IN COMBINATION: A SUBSTANTIALLY VERTICALLY DISPOSED COLUMN PHASE TO AN UPPER TUBE, AND A SEPARATION SUBSTANCE; A PRECISION PUMP DELIVERING A MOBILE PHASE TO AN UPPER PART OF SAID COLUMN; A PISTON IN SAID PUMP AND A FIRST DRIVING MEANS RECIPROCATING THE PISTON; AN EXCHANGEABLE COLLECTION VESSEL ADAPTED TO RECEIVE A FRACTION OF ELUATE ISSUING FROM A LOWER PART OF SAID COLUMN; SIGNALING MEANS INDICATING A DESIRED REPLACEMENT OF SAID COLLECTING BY ANOTHER COLLECTING VESSEL TO RECEIVE A FURTHER FRACTION OF ELUATE; ELECTROMECHANICAL MEANS OPERATING THE SIGNALING MEANS IN SYNCHRONOUS RELATION TO A PREDETERMINED SUCCESSION OF STROKES OF THE PUMP PISTON; AND SECOND DRIVING MEANS FOR SAID ELECTROMECHANICAL MEANS OPERATIVELY CONNECTING SAID ELECTROMECHANICAL MEANS WITH SAID FIRST DRIVING MEANS. 